The present invention relates to polypeptide compounds which possess antagonist properties against bombesin, or bombesin-like peptides, and are useful in the treatment of disease, particularly human small-cell lung cancer. Zollinger-Ellison syndrome or pancreatic cancer. The invention thus provides the polypeptides, processes for preparing them, pharmaceutical compositions containing them, and their use in medicine.
Bombesin is a tetradecapeptide originally isolated from the skin of a frog. It has the formula EQU pGlu-Gln-Arg-Leu-Gly-Asn-Gln-Trp-Ala-Val-Gly-His-Leu-Met-NH.sub.2 (SEQ ID NO: 1)
Gastrin releasing peptide is a 27 amino acid peptide isolated from the porcine gut. The last ten amino acids at the C-terminus of gastrin releasing peptide correspond with one amino acid alteration (3) to the last ten amino acids of bombesin, viz: EQU H-Gly-Asn-His-Trp-Ala-Val-Gly-His-Leu-Met-NH.sub.2 (SEQ ID NO: 2)
It has been reported (J. H. Walsh and J. R. Reeve, Peptides 6, (3), 63-68, (1985)) that bombesin and bombesin-like peptides such as gastrin releasing peptide are secreted by human small-cell lung cancer (SCLC) cells. It has been postulated (P. J. Woll and E. Rozengurt, PNAS 85, 1859-1863, (1988)) that gastrin releasing factor antagonists would bind competitively to bombesin receptors in animals and would therefore be of use in the treatment of SCLC and/or in the control of clinical symptoms associated with this disease and due to hypersecretion of this peptide hormone. Analogues of bombesin have been shown to inhibit the binding of gastrin releasing peptide to a SCLC cell line and to inhibit the growth of SCLC cells in-vitro and in-vivo (S. Mahmoud et al., Cancer Research, 51, 1798-1802 (1991).
Several bombesin antagonists have been disclosed, for example [Leu.sup.13 -.psi.(CH.sub.2 --NH)-Leu.sup.14 ] bombesin and [Ala.sup.9 -.psi.(CH.sub.2 --NH)-Val.sup.10 Leu.sup.14 ] bombesin (Coy et al, J. Biol Chem., 1988, 263, 5056) and
4-Pyridyl-CO-His-Trp-Ala-Val- D-Ala-His-Leu-OMe, PA0 4-Pyridyl-CO-His-Trp-Ala-Val- D-Ala-His-Leu-NHMe, PA0 4-Pyridyl-CO-His-Trp-Ala-Val- D-Ala-His-MeLeu-OMe, PA0 3-Pyridyl-CO-His-Trp-Ala-Val- D-Ala-His-MeLeu-OMe, PA0 4-Pyridyl-CO-His-Trp-Ala-Val- D-Ala-Lys(Z)-MeLeu-OMe, PA0 3-Indolyl-Co-His-Trp-Ala-Val- D-Ala-His-Leu-OMe, PA0 4-Pyridyl-CO-His-Trp-Ala-Val- D-Ala-His-MeLeu-NHMe, PA0 4-Pyridyl-CO-His-Trp-Ala-Val- D-Ala-Lys(Z)-Leu-NHMe, PA0 4-Pyridyl-CO-His-Trp-Ala-Val- D-Ala-Lys(COCH.sub.2 Ph)-Leu-NHMe and PA0 4-Pyridyl-CO-His-Trp-Ala-Val- D-Ala-Lys(COCH.sub.2 CH.sub.2 Ph)-Leu-NHMe. PA0 (European Patent Application No. 345990A). PA0 .psi.=psi(CH.sub.2 NH) PA0 Ada=1-adamantanecarboxylic acid PA0 CPenc=aminocyclopentanecarboxylic acid PA0 Mox=methoxinine PA0 Des NH.sub.2 Pro=1-cyclopentanecarboxylic acid PA0 Des NH.sub.2 Tyr=(4'-hydroxy)-3-phenylpropionic acid PA0 ThiAla=3-(2-thienyl)-alanine PA0 D-tBuGly= D-tertiary-butyl-glycine (tertiary- D-leucine) PA0 Suitably A is phenyl, naphthyl, phenothiazinyl or indolyl. Preferably A is phenyl or naphthyl. PA0 Suitable substituents for the aromatic ring Ar include hydroxy, phenyl, halo, C.sub.1-4 alkyl or C.sub.1-4 alkoxy optionally substituted by halo. PA0 Preferably n is 2. PA0 Suitably X terminates in a des NH.sub.2 moiety. PA0 Suitably X.sup.8 is des NH.sub.2 Tyr or des NH.sub.2 Pro. Suitably X.sup.9 is Gly or D-Ala. PA0 Suitably X.sup.10 is D-Phe. Preferably X is des NH.sub.2 Phe, des NH.sub.2 Tyr, des NH.sub.2 TyrPro (or D-Pro) Arg (or D-Arg). PA0 N-((R)-2-(6-Methoxy-2-Naphthyl)Propionyl)-HisTrpAlaVal D-AlaHis D-Pro.psi.Nle-NH.sub.2 PA0 N-((S)-2-(6-Methoxy-2-Naphthyl)Propionyl)-HisTrpAlaVal D-AlaHis D-Pro.psi.Nle-NH.sub.2 PA0 N-((S)-3-Phenylbutyryl)-HisTrpAlaVal D-AlaHis D-Pro.psi.Nle-NH.sub.2 PA0 N-((R)-3-Phenylbutyryl)-HisTrpAlaVal D-AlaHis D-Pro.psi.Nle-NH.sub.2 PA0 N-((3-Phenyl)Propionyl)-HisTrpAlaVal D-Ala(3-(2-Thi)-Ala) D-Pro.psi.Nle-NH.sub.2 PA0 N-((S)-3,3,3-Trifluoro-2-Methoxy-2-Phenyl-Propionyl)-HisTrpAlaVal D-Pro.psi.Nle-NH.sub.2 PA0 N-((R)-3,3,3-Trifluoro-2-Methoxy-2-Phenyl-Propionyl)-HisTrpAlaVal D-Pro.psi.Nle-NH.sub.2 PA0 N-3-(((4'-Hydroxy)Phenyl)Propionyl)-Pro D-ArgGly D-PheHisTrpAlaValGlyHis D-Pro.psi.Nle-NH.sub.2 PA0 N-(((4'Hydroxy)-3-Phenyl)Propionyl)-Pro D-ArgHisTrpAlaVal D-AlaHis D-Pro.psi.Leu-NH.sub.2 PA0 N-((3-Phenyl)Propionyl)-HisTrpAlaVal D-AlaHis D-Pro.psi.mox-NH.sub.2 PA0 N-((3-Phenyl)Propionyl)-HisTrpAlaVal D-Pro.psi.Phe-NH.sub.2 PA0 N-((3-Phenyl)Propionyl)-TrpAlaVal D-AlaHis D-Pro.psi.Leu-NH.sub.2 PA0 N-((3-Phenyl)Propionyl)-HisTrpProVal D-ProHis D-Pro.psi.Leu-NH.sub.2 PA0 N-3-(((3'-Trifluoromethyl)Phenyl)Propionyl)-HisTrpAlaVal D-AlaHis D-Pro.psi.Leu-NH.sub.2 PA0 N-((3-Phenyl)Propionyl)-(3-(2-Thi)-Ala)TrpAlaVal D-AlaHis D-Pro.psi.Leu-NH.sub.2 PA0 N-((deamino-Pro)- D-Arg D-Ala D-PheHisTrpAlaValGlyHis D-Pro.psi.Nle-NH.sub.2 PA0 N-((3-Phenyl)Propionyl)-HisTrpAlaValGlyHis D-Pro.psi.Nle-NH.sub.2 PA0 N-((deamino-Pro)- D-Arg D-Ala D-PheHisTrpAlaVal D-AlaHis D-Pro.psi.Nle-NH.sub.2 PA0 N-((3-Phenyl)Propionyl)-HisTrpAlaVal D-AlaHis D-Pro.psi.Nle-NH.sub.2 PA0 TyrPro D-ArgGly D-PheHisTrpAlaValGlyHis D-Pro.psi.Nle-NH.sub.2 PA0 D-ArgGly D-PheHisTrpAlaValGlyHis D-Pro.psi.Nle-NH.sub.2 PA0 N-((3-Phenyl)Propionyl)-HisTrpAlaVal D-AlaHis D-ProPhe-NH.sub.2 PA0 N-((3-Phenyl)Propionyl)-HisTrpAlaVal D-AlaHis D-Pro.psi.(3-(2-Naphthyl)- D-Ala)-NH.sub.2 PA0 N-((3-Phenyl)Propionyl)-HisTrpAlaVal D-PheHis D-Pro.psi.Phe-NH.sub.2 PA0 D-PheHisTrpAlaVal D-AlaHis D-Pro.psi.Phe-NH.sub.2 PA0 N-((3-Phenyl)Propionyl)- D-ProArgGly D-PheHisTrpAlaVal D-AlaHis D-Pro.psi.Phe-NH.sub.2 PA0 N-((3-Phenyl)Propionyl)-(3-(2-Thi)-Ala)-TrpAlaVal D-AlaHis D-Pro.psi.Phe-NH.sub.2 PA0 N-((3-Phenyl)Propionyl)-HisTrpAlaVal-(Sarcosine)-His D-Pro.psi.Phe-NH.sub.2 PA0 N-3-(((4'-Hydroxy)Phenyl)Propionyl)-HisTrpAlaVal D-AlaHis D-Pro.psi.Phe-NH.sub.2 PA0 N-(((2',6'-Dichloro)-2-Phenyl)Acetyl)-HisTrpAlaVal D-AlaHis D-Pro.psi.Nle-NH.sub.2 PA0 N(((3',4'-Dichloro)-2-Phenyl)Acetyl)-HisTrpAlaValD-AlaHis D-Pro.psi.Nle-NH.sub.2 PA0 N-(((4'-Hydroxy)-2-Phenyl)Acetyl)-HisTrpAlaVal D-AlaHis D-Pro.psi.Nle-NH.sub.2 PA0 N-(1-Naphthoyl)-HisTrpAlaVal D-AlaHis D-Pro.psi.Nle-NH.sub.2 PA0 N-((3,7-Dihydroxy)-2-Naphthoyl)-HisTrpAlaVal D-AlaHis D-Pro.psi.Nle-NH.sub.2 PA0 N-((3,4-Dihydroxy)-2-Phenyl)Acetyl)-HisTrpAlaVal D-AlaHis D-Pro.psi.Nle-NH.sub.2 PA0 N-(2-(3-Pyridyl)Acetyl)-HisTrpAlaVal D-AlaHis D-Pro.psi.Nle-NH.sub.2 PA0 N-(2-(2-Thienyl)Acetyl)-HisTrpAlaVal D-AlaHis D-Pro.psi.Nle-NH.sub.2 PA0 N-(((3-Fluoro)-3-Phenyl)Prop-ionyl)-HisTrpAlaVal D-AlaHis D-Pro.psi.Nle-NH.sub.2 PA0 N-(((4-hydroxy-3-methoxy)-2-Phenyl)Acetyl)-HisTrpAlaVal D-AlaHis D-Pro.psi.Nle-NH.sub.2 PA0 N-(((R)-(-)-2-Phenyl)Propionyl)-HisTrpAlaVal D-AlaHis D-Pro.psi.Phe-NH.sub.2 PA0 N-(((S)-(+)-2-Phenyl)Propionyl)-HisTrpAlaVal D-AlaHis D-Pro.psi.Phe-NH.sub.2 PA0 N-(((Trans)-2-Phenyl)-Cyclopro-panoyl)-HisTrpAlaVal D-AlaHis D-Pro.psi.Phe-NH.sub.2 PA0 N-(3-(10-Phenothiazinyl)Propionyl-HisTrpAlaVal D-AlaHis D-Pro.psi.Phe-NH.sub.2 PA0 N-((3-Methyl-3-Phenyl)Butyryl)-HisTrpAlaVal D-AlaHis D-Pro.psi.Phe-NH.sub.2 PA0 N-(((2'-Trifluoromethyl)-2-Phenyl)Acetyl)-HisTrpAlaVal D-AlaHis D-Pro.psi.Phe-NH.sub.2 PA0 N-(((3'-Trifluoromethyl)-2-Phenyl)Acetyl)-HisTrpAlaVal D-AlaHis D-Pro.psi.Phe-NH.sub.2 PA0 N-(((4'-Trifluoromethyl)-2-Phenyl)Acetyl)-HisTrpAlaVal D-AlaHis D-Pro.psi.Phe-NH.sub.2 PA0 N-(((2',3'-Difluoro)-2-Phenyl)Acetyl)-HisTrpAlaVal D-AlaHis D-Pro.psi.Phe-NH.sub.2 PA0 N-(((2',4'-Difluoro)-2-Phenyl)Acetyl)-HisTrpAlaVal D-AlaHis D-Pro.psi.Phe-NH.sub.2 PA0 N-(((2',6'-Difluoro)-2-Phenyl)Acetyl)-HisTrpAlaVal D-AlaHis D-Pro.psi.Phe-NH.sub.2 PA0 N-(((2-Amino)-2-Phenyl)Acetyl)-HisTrpAlaVal D-AlaHis D-Pro.psi.Phe-NH.sub.2 PA0 N-(1-Naphthoyl)-HisTrpAlaVal D-AlaHis D-Pro.psi.Phe-NH.sub.2 PA0 N-(((3',4',5'-Trimethoxy)-3-Phenyl)Propionyl)-HisTrpAlaVal D-AlaHis D-Pro.psi.Phe-NH.sub.2 PA0 N-((6'-Methoxy)-2-(2-Naphthoyl)Propionyl)-HisTrpAlaVal D-AlaHis D-Pro.psi.Phe-NH.sub.2 PA0 N-(((3'-Trifluoromethyl)-3-Phenyl)Propionyl)-HisTrpAlaVal D-AlaHis D-Pro.psi.Phe-NH.sub.2 PA0 N-(((S)-3-Phenyl)Butyryl)-HisTrpAlaVal D-AlaHis D-Pro.psi.Phe-NH.sub.2 PA0 N-(((4'-Methoxy)-3-Phenyl)Propionyl)-HisTrpAlaVal D-AlaHis D-Pro.psi.Phe-NH.sub.2 PA0 N-((((S)-2-Hydroxy)-2-Phenyl)Acetyl)-HisTrpAlaVal D-AlaHis D-Pro.psi.Phe-NH.sub.2 PA0 N-((3-Phenyl)Propionyl)-HisTrpAlaVal D-AlaHisPro.psi.Phe-NH.sub.2 PA0 N-((2-Methyl-2-Phenyl)Propionyl)-HisTrpAlaVal D-AlaHis D-Pro.psi.Phe-NH.sub.2 PA0 N-(3-(1-Naphthyl)Propionyl)-HisTrpAlaVal D-AlaHis D-Pro.psi.Phe-NH.sub.2 PA0 N-(((R)-3-Phenyl)Butyryl)-HisTrpAlaVal D-AlaHis D-Pro.psi.Phe-NH.sub.2 PA0 N-((9-Fluoroenoyl)1-Carbonyl)-HisTrpAlaVal D-AlaHis D-Pro.psi.Phe-NH.sub.2 PA0 N-(((2'-Methoxy)-3-Phenyl)Propionyl)-HisTrpAlaVal D-AlaHis D-Pro.psi.Phe-NH.sub.2 PA0 N-(((2',5'-Dimethoxy)-3-Phenyl)Propionyl)-HisTrpAlaVal D-AlaHis D-Pro.psi.Phe-NH.sub.2 PA0 N-((3-Phenyl)Propionyl)-HisTrpAlaVal D-AlaHis D-Pro.psi.Tyr-NH.sub.2 PA0 N-(((2',3'-Dimethoxy)-3-Phenyl)Propionyl)-HisTrpAlaVal D-AlaHis D-Pro.psi.Phe-NH.sub.2 PA0 N-((3-Phenyl)Propionyl)-HisTrpAlaVal D-AlaHis(3-(2-Pyrrolidinyl-3-Hydroxy)Propionyl)-Phe-NH.sub.2 PA0 ((Isoquinolylcarbonyl)-HisTrpAlaVal D-AlaHis D-Pro.psi.Phe-NH.sub.2 PA0 N-((3-Phenyl)Propionyl)-HisTrpAlaVal D-AlaHis D-Pro.psi.Phe-NH.sub.2 PA0 (a) the removal of one or more conventional peptide protecting groups from a protected polypeptide to give a polypeptide of the invention of formula I; PA0 (b) the formation of an amide bond by coupling two peptide units, one containing a carboxylic acid group, or a reactive derivative thereof, and the other containing an amino group, such that a protected or unprotected polypeptide having the sequence indicated in formula I is produced whereafter, if necessary, the protecting groups are removed using process (a) above. PA0 (a) polypeptides of the invention for use in therapy; PA0 (b) pharmaceutical formulations containing polypeptides of the invention, at least one pharmaceutical carrier or excipient and, optionally, one or more other therapeutic ingredients; PA0 (c) the use of polypeptides of the invention in the manufacture of a medicament for the treatment of cancer; PA0 (d) the use of a polypeptide of the invention in the manufacture of a medicament for the treatment of: PA0 (e) a method for inhibiting the growth of cells that are sensitive to the growth promoting activity of gastrin releasing peptide in a mammal (such as a human) in need of such treatment which comprises the administration to said mammal of a growth inhibiting amount of a polypeptide of the invention to said mammal.
In the formula (I) below and throughout this specification, the amino acid residues are designated by their standard abbreviations (Pure and Applied Chemistry, 1974, 40, 317-331; European Journal of Biochemistry, 1984, 138, 9-37).
For the avoidance of doubt it is stated that: amino acid symbols denote the L-configuration unless otherwise indicated by D or DL appearing before the symbol and separated from it by a hyphen. (R) and (S) are the standard designations for molecular configuration.
The following abbreviations are used:
When the Ada,CPenc,Mox,des NH.sub.2 Pro,des NH.sub.2 Tyr groups are in a polypeptide chain they are in the carbonyl form.
It has now been discovered that a further group of polypeptides have potent bombesin antagonist activity.
The compounds of the present invention inhibit the production of gastrin releasing peptide in mammalian cells and are therefore useful in controlling the clinical symptoms of diseases which cause the secretion of hypersecretion of gastrin releasing peptide (e.g., SCLC).